A Lab‐Built Respirometer for Plant and Animal Cell Culture

Lamboursain, Laurence; St‐Onge, Frédéric; Jolicœur, Mario

doi:10.1021/bp015511j

Cited by 20 publications

(12 citation statements)

References 33 publications

(41 reference statements)

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“…OD 600 measurements and cell counts were performed immediately before putting cells in the oxygen sensing setup. The oxygen depletion rate was monitored by using a lab-built respirometer comprising a polarographic dissolved oxygen probe in a glass syringe with constant agitation (26). Oxygen consumption rates were expressed as percent O 2 per minute per 1 ϫ 10 6 cells.…”

Section: Methodsmentioning

confidence: 99%

Increased Respiration in the sch9Δ Mutant Is Required for Increasing Chronological Life Span but Not Replicative Life Span

Lavoie

Whiteway

2008

Eukaryot Cell

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Loss of the protein kinase Sch9p increases both the chronological life span (CLS) and the replicative life span (RLS) of Saccharomyces cerevisiae by mimicking calorie restriction, but the physiological consequences of SCH9 deletion are poorly understood. By transcriptional profiling of an sch9Delta mutant, we show that mitochondrial electron transport chain genes are upregulated. Accordingly, protein levels of electron transport chain subunits are increased and the oxygen consumption rate is enhanced in the sch9Delta mutant. Deletion of HAP4 and CYT1, both of which are essential for respiration, revert the sch9Delta mutant respiratory rate back to a lower-than-wild-type level. These alterations of the electron transport chain almost completely blocked CLS extension by the sch9Delta mutation but had a minor impact on the RLS. SCH9 thus negatively regulates the CLS and RLS through inhibition of respiratory genes, but a large part of its action on life span seems to be respiration independent and might involve increased resistance to stress. Considering that TOR1 deletion also increases respiration and that Sch9p is a direct target of TOR signaling, we propose that SCH9 is one of the major effectors of TOR repression of respiratory activity in glucose grown cells.

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Section: Methodsmentioning

confidence: 99%

Increased Respiration in the sch9Δ Mutant Is Required for Increasing Chronological Life Span but Not Replicative Life Span

Lavoie

Whiteway

2008

Eukaryot Cell

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“…Plant cell calli were obtained as described in Lamboursain et al (2002). The cell suspension was sub-cultured when the packed cell volume after 5 min of sedimentation reached 70%-80% of the total volume (i.e., every 10-15 days).…”

Section: Plant Cell Line Culture Conditions and Media Compositionmentioning

confidence: 99%

“…The oxygen uptake rate (OUR) of the cells was assessed in a custom built respirometer as described by Lamboursain et al (2002).…”

Section: Respirometric Measurementsmentioning

confidence: 99%

Critical influence of Eschscholzia californica cells nutritional state on secondary metabolite production

Lamboursain

Jolicœur

2005

Biotech & Bioengineering

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In this study, we investigated the influence of initial internal nutrient concentrations at the time of elicitation on the ability of Eschscholzia californica (EC) cells to produce alkaloids. Three EC cell suspensions cultivated in culture media differing in their PO4(3-) and NO3- contents were sampled daily for 12 days and analyzed for extracellular and intracellular nutrient concentrations. The ability of the cells to produce alkaloids was tested along the three cell suspension cultures. Sampled cells were then further cultured for 7 days in a production medium containing the elicitor and an extraction resin. The alkaloid production of the cells was measured 7 days post-elicitation. In the low-N medium, starch, glucose, and phosphate contents in the biomass was increased by 470, 1624 and 70%, respectively, 10 days after inoculation compared to the control culture in standard B5 medium. Cell concentration was significantly reduced from 10.3 to 8.6 millions cell/mL on this low-N medium compared to the control, nevertheless alkaloid production was multiplied by 39 at day 10 when cells were elicited. Cells grown on the low-N or low-P media accumulated 83% and 188% more carbon, respectively, than control cells at the end of the culture. This intracellular C was mainly stored in the form of starch in low-P medium and both in the form of starch and glucose in the low-N medium. The ability of EC cells to produce alkaloids upon elicitation was shown to be strongly dependent on the initial intracellular C and P content at the time of elicitation. This suggests that reproducibility and productivity during EC cell culture could be enhanced by manipulating the intracellular C and P content at the time of elicitation.

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“…Respirometry has been used extensively for this purpose, and also to monitor shoot and/or root O 2 flux (Lamboursain et al 2002;De Dobbeleer et al 2006;Cloutier et al 2009;Gupta et al 2009). Microassays have also been used to analyze cell/tissue respiration by fluorescence intensity measured in microtiter plates (O'Riordan et al 2000;Alderman et al 2004;Kupper et al 2004;Serrano et al 2007;Zitova et al 2009).…”

Section: Introductionmentioning

confidence: 99%

Self-referencing optrodes for measuring spatially resolved, real-time metabolic oxygen flux in plant systems

McLamore

Jaroch

Chatni

et al. 2010

Planta

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The ability to non-invasively measure metabolic oxygen flux is a very important tool for physiologists interested in a variety of questions ranging from basic metabolism, growth/development, and stress adaptation. Technologies for measuring oxygen concentration near the surface of cells/tissues include electrochemical and optical techniques. A wealth of knowledge was gained using these tools for quantifying real-time physiology. Fiber-optic microprobes (optrodes) have recently been developed for measuring oxygen in a variety of biomedical and environmental applications. We have adopted the use of these optical microsensors for plant physiology applications, and used the microsensors in an advanced sensing modality known as self-referencing. Self-referencing is a non-invasive microsensor technique used for measuring real-time flux of analytes. This paper demonstrates the use of optical microsensors for non-invasively measuring rhizosphere oxygen flux associated with respiration in plant roots, as well as boundary layer oxygen flux in phytoplankton mats. Highly sensitive/selective optrodes had little to no hysteresis/calibration drift during experimentation, and an extremely high signal-to-noise ratio. We have used this new tool to compare various aspects of rhizosphere oxygen flux for roots of Glycine max, Zea mays, and Phaseolus vulgaris, and also mapped developmentally relevant profiles and distinct temporal patterns. We also characterized real-time respiratory patterns during inhibition of cytochrome and alternative oxidase pathways via pharmacology. Boundary layer oxygen flux was also measured for a phytoplankton mat during dark:light cycling and exposure to pharamacological inhibitors. This highly sensitive technology enables non-invasive study of oxygen transport in plant systems under physiologically relevant conditions.

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A Lab‐Built Respirometer for Plant and Animal Cell Culture

Cited by 20 publications

References 33 publications

Increased Respiration in the sch9Δ Mutant Is Required for Increasing Chronological Life Span but Not Replicative Life Span

Increased Respiration in the sch9Δ Mutant Is Required for Increasing Chronological Life Span but Not Replicative Life Span

Critical influence of Eschscholzia californica cells nutritional state on secondary metabolite production

Self-referencing optrodes for measuring spatially resolved, real-time metabolic oxygen flux in plant systems

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